Development and validation of a bioanalytical method for quantification of LNA-i-miR-221, a 13-mer oligonucleotide, in rat plasma using LC-MS/MS
Development and validation of a bioanalytical method for quantification of LNA-i-miR-221, a 13-mer oligonucleotide, in rat plasma using LC-MS/MS
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DOI：
10.1016/j.jpba.2017.12.027
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发表时间：
2018-02-20
影响因子：
3.4
通讯作者：
Breda, M.
中科院分区：
文献类型：
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作者：
Franzoni, S.;Vezzelli, A.;Breda, M.
LNA-i-miR-221 是一种 13 聚体寡核苷酸，是一种新型 miR-221 抑制剂，可用作治疗多发性骨髓瘤的新药。在此，开发并验证了离子对反相液相色谱-串联质谱 (LC-MS/MS) 方法，用于定量大鼠血浆中的 LNA-i-miR-221。血浆样品首先采用苯酚/氯仿/异戊醇液-液萃取，然后进行固相萃取。使用六氟-2-丙醇/三乙胺缓冲液和甲醇作为流动相，在HALO C18柱上通过梯度系统以0.4毫升/分钟的流速进行色谱分离。在这些条件下，LNA-i-miR-221 和类似物内标在 1.2 分钟时共洗脱。使用负电喷雾电离 (ESI) 接口以多反应监测 (MRM) 模式进行检测。该测定在 10-10000 ng/mL 的校准范围内显示出良好的线性。精密度、准确度和回收率值分别为
LNA-i-miR-221, a 13-mer oligonucleotide, is a new miR-221 inhibitor that could be used as a novel drug for multiple myeloma. Herein, an ion-pair reversed phase liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the quantification of LNA-i-miR-221 in rat plasma. Plasma samples were prepared with an initial phenol/chloroform/isoamyl alcohol liquid-liquid extraction followed by a solid phase extraction. Chromatographic separation was performed with a gradient system on a HALO C18 column using hexafluoro-2-propanol/triethylamine buffer and methanol as mobile phase at a flow rate of 0.4 mi./min. Under these conditions LNA-i-miR-221 and the analogue internal standard are co-eluted at 1.2 min. The detection was carried out in multiple reaction monitoring (MRM) mode using a negative electrospray ionization (ESI) interface. The assay showed a good linearity within the calibration range 10-10000 ng/mL. The precision, accuracy, and recovery values were found to be